Unique Human and Mouse β-cell Senescence-Associated Secretory Phenotype (SASP) Reveal Conserved Signaling Pathways and Heterogeneous Factors

Midha, Ayush; Hui, Pei; Abarca, Cristian; Andle, Joshua; Carapeto, Priscila; Bonner-Weir, Susan; Aguayo-Mazzucato, Cristina

doi:10.2337/figshare.14125511

Cited by 6 publications

(11 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We next assessed whether CDK inhibitors p21 and/or p16 Ink4a were upregulated at later stages after drug removal in NIT1 cells ( Figure 2 ). Notably, at the 72 h post-treatment timepoint, Cdkn1a but not Cdkn2a was upregulated in MIN6 cells and this persisted into late stage senescence at 14 days post-drug removal ( Figure 2A ), consistent with previous results [26]. Similarly, NIT1 cells also upregulated Cdkn1a expression at 72 h post-treatment and 7 days post-drug removal and also showed no upregulation of Cdkn2a ( Figure 2B ).…”

Section: Resultssupporting

confidence: 90%

“…Luminex assays for SASP factors IL-6, Serpine1 and Igfbp3 [9] on conditioned media showed dramatically increased secretion of Serpine1 and Igfbp3, but not IL-6 from senescent NIT1 cells as compared with controls whereas none of these factors were secreted by senescent MIN6 cells ( Figure 3C and 3D ). In contrast, senescent MIN6 cells secreted Ccl11 and Ccl4 ( Supplementary Figure 4A ) consistent with previous reports [26,35]. Given that EV secretion is increased in senescent cells with DNA damage [36] we also assayed for total EV secretion from senescent NIT1 cells.…”

Section: Resultssupporting

confidence: 84%

“…To develop a mouse beta cell culture model for senescence in T1D, we investigated senescent phenotypes in the NIT1 cell line, derived from a beta cell tumor in the T1D susceptible NOD/LtJ mouse strain [17] in response to DNA double-strand break damage. For comparison, we also examined the MIN6 beta cell line derived from the non-T1D susceptible C57BL6 strain [16], which was shown to develop senescent phenotypes in response to DNA damage with bleomycin [26]. Both lines have been immortalized via the expression of a rat insulin promoter driving the Simian virus 40 large T antigen (SV40 TAg).…”

Section: Resultsmentioning

confidence: 99%

See 2 more Smart Citations

DNA damage models phenotypes of β cell senescence in Type 1 Diabetes

Brawerman

Thompson

2021

Preprint

View full text Add to dashboard Cite

Pancreatic beta cells are essential endocrine cells in the islets of Langerhans that respond to increases in blood glucose by secreting insulin and their dysfunction and death drives the development of diabetes. Beta cell senescence involving a DNA damage response (DDR) was recently shown to contribute to the pathogenesis of Type 1 Diabetes (T1D), however, a basic mechanistic understanding of how senescence develops in beta cells is lacking. Here, we investigated senescent phenotypes arising in the mouse beta cell line NIT1 derived from the T1D susceptible nonobese diabetic (NOD) mouse strain and the transcriptome response in human islets after induction of DNA damage. Sub-lethal DNA damage in NIT1 cells led to several classical hallmarks of senescence including the DDR, growth arrest, enlarged flattened morphology and a senescence-associated secretory phenotype (SASP) resembling what occurs during T1D. RNA-seq analysis on human islets with DNA double-strand break damage revealed a coordinated p53-p21 transcriptional response and upregulation of genes involved in prosurvival signaling and SASP. Taken together, these findings suggest that some of the phenotypes of mouse and human beta cell senescence during T1D can be induced by DNA damage in NIT1 cells and human islets in culture.

show abstract

Section: Resultssupporting

confidence: 90%

Section: Resultssupporting

confidence: 84%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

DNA damage models phenotypes of β cell senescence in Type 1 Diabetes

Brawerman

Thompson

2021

Preprint

View full text Add to dashboard Cite

show abstract

“…Further, we address a major knowledge gap: β cells are a crucial endocrine cell type, yet little is known about the β cell secretome, especially in humans. Few studies reported secretome composition of rodent insulinoma cell lines (20)(21)(22), while the secretome of human immune-cell depleted islet tissue has been profiled using the SOMAscan 1,300 proteomics assay only this year (23). The SOMAscan study suggests that the human and murine secretomes may be quite distinct (23), making it unclear to what extent findings of secretome studies in murine β cells can be extrapolated to human.…”

Section: Validity Of Endoc-βh1 Cells As a Human β Cell Model Has Been Previously Established Based Onmentioning

confidence: 99%

“…Few studies reported secretome composition of rodent insulinoma cell lines (20)(21)(22), while the secretome of human immune-cell depleted islet tissue has been profiled using the SOMAscan 1,300 proteomics assay only this year (23). The SOMAscan study suggests that the human and murine secretomes may be quite distinct (23), making it unclear to what extent findings of secretome studies in murine β cells can be extrapolated to human. In this study, we present the first characterization of human β cell line EndoC-βH1 secretome by Data Independent Acquisition (DIA) proteomics.…”

Section: Validity Of Endoc-βh1 Cells As a Human β Cell Model Has Been Previously Established Based Onmentioning

confidence: 99%

Characterization of the secretome, transcriptome and proteome of human β cell line EndoC-βH1

Ryaboshapkina

Saitoski

Hamza

et al. 2021

Preprint

View full text Add to dashboard Cite

Early diabetes research is hampered by limited availability, variable quality and instability of human pancreatic islets in culture. Little is known about the human β cell secretome, and recent studies question translatability of rodent β cell secretory profiles. Here, we verify representativeness of EndoC-βH1, one of the most widely used human β cell lines, as a translational human β cell model based on omics and characterize the EndoC-βH1 secretome. We profiled EndoC-βH1 cells using RNA-seq, Data Independent Acquisition (DIA) and Tandem Mass Tag proteomics of cell lysate. Omics profiles of EndoC-βH1 cells were compared to human β cells and insulinomas. Secretome composition was assessed by DIA proteomics. Agreement between EndoC-βH1 cells and primary adult human β cells was ~90% for global omics profiles as well as for β cell markers, transcription factors and enzymes. Discrepancies in expression were due to elevated proliferation rate of EndoC-βH1 cells compared to adult β cells. Consistently, similarity was slightly higher with benign non-metastatic insulinomas. EndoC-βH1 secreted 671 proteins in untreated baseline state and 3,278 proteins when stressed with non-targeting control siRNA, including known β cell hormones INS, IAPP, and IGF2. Further, EndoC-βH1 secreted proteins known to generate bioactive peptides such as granins and enzymes required for production of bioactive peptides. Unexpectedly, exosomes appeared to be a major mode of secretion in EndoC-βH1 cells. We believe that secretion of exosomes and bioactive peptides warrant further investigation with specialized proteomics workflows in future studies.

show abstract

IL-1beta promotes the age-associated decline of beta cell function

et al. 2021

View full text Add to dashboard Cite

Aging is the prime risk factor for the development of type 2 diabetes. We investigated the role of the interleukin-1 (IL-1) system on insulin secretion in aged mice. During aging, expression of the protective IL-1 receptor antagonist decreased in islets, whereas IL-1beta gene expression increased specifically in the CD45 + islet immune cell fraction. One-year-old mice with a whole-body knockout of IL-1beta had higher insulin secretion in vivo and in isolated islets, along with enhanced proliferation marker Ki67 and elevated size and number of islets. Myeloid cell-specific IL-1beta knockout preserved glucose-stimulated insulin secretion during aging, whereas it declined in control mice. Isolated islets from aged myeloIL-1beta ko mice secreted more insulin along with increased expression of Ins2, Kir6.2, and of the cell-cycle gene E2f1. IL-1beta treatment of isolated islets reduced E2f1, Ins2, and Kir6.2 expression in beta cells. We conclude that IL-1beta contributes the age-associated decline of beta cell function.

show abstract

Unique Human and Mouse β-cell Senescence-Associated Secretory Phenotype (SASP) Reveal Conserved Signaling Pathways and Heterogeneous Factors

Cited by 6 publications

References 0 publications

DNA damage models phenotypes of β cell senescence in Type 1 Diabetes

DNA damage models phenotypes of β cell senescence in Type 1 Diabetes

Characterization of the secretome, transcriptome and proteome of human β cell line EndoC-βH1

IL-1beta promotes the age-associated decline of beta cell function

Contact Info

Product

Resources

About